Literature summary for 3.2.1.132 extracted from

Lacombe-Harvey, M.E.; Fukamizo, T.; Gagnon, J.; Ghinet, M.G.; Dennhart, N.; Letzel, T.; Brzezinski, R.
Accessory active site residues of Streptomyces sp. N174 chitosanase: variations on a common theme in the lysozyme superfamily (2009), FEBS J., 276, 857-869.

Search for more literature for 3.2.1.132

Cloned(Commentary)

Cloned (Comment)	Organism
expression of CsnN174 in Escherichia coli JM109. T45S chitosanase mutant introduced into Streptomyces lividans	Streptomyces sp. N174

Protein Variants

Protein Variants	Comment	Organism
D40E	lower activity than mutant D40G	Streptomyces sp. N174
D40G	reaction time is 20 min. It is distinguished from wild-type by a lower activity, without changing the mechanism of hydrolysis or the mode of interaction with substrate	Streptomyces sp. N174
D40G/T45D	reaction time is 100 min. It has 0.1% of wild-type specific activity when tested on chitosan substrate	Streptomyces sp. N174
D40G/T45E	has 0.03% of wild-type specific activity when tested on chitosan substrate	Streptomyces sp. N174
D40N	lower activity than mutant D40G	Streptomyces sp. N174
E36A	reaction time is 10 min	Streptomyces sp. N174
E36A/D40G	reaction time is 100 min. Kcat is ca. 18times lower than that of the single mutant D40G. Rate of (GlcN)6 degradation is enhanced by sodium azide	Streptomyces sp. N174
E36D	reaction time is 10 min	Streptomyces sp. N174
E36N	reaction time is 10 min	Streptomyces sp. N174
E36Q	reaction time is 10 min	Streptomyces sp. N174
E36Q/D40G	reaction time is 100 min. Kcat is more than 5times lower than that of the single mutant D40G	Streptomyces sp. N174
T45E	has a very low residual activity. Activity of this mutant can not be enhanced by sodium azide	Streptomyces sp. N174
T45H	results in a complete loss of activity	Streptomyces sp. N174
T45S	reaction time is 20 min. It is quite active, keeps ca. 71% of the specific activity of the wild-type enzyme	Streptomyces sp. N174
V148T	has 10% of wild-type activity when tested on chitosan substrate	Streptomyces sp. N174

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	extent of growth inhibition is dependent on chitosan concentration, average molecular mass, the pH of the medium and salt composition. Growth inhibition can be suppressed by the expression of CsnN174 in Escherichia coli JM109	Streptomyces sp. N174

Organism

Organism	UniProt	Comment	Textmining
Streptomyces sp. N174	P33665	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.01	-	mutant T45H	Streptomyces sp. N174
0.011	-	mutant T45E	Streptomyces sp. N174
0.02	-	mutant D40G/T45E	Streptomyces sp. N174
0.06	-	mutant D40G/T45D	Streptomyces sp. N174
0.07	-	mutant E36A/D40G	Streptomyces sp. N174
0.3	-	mutant E36Q/D40G	Streptomyces sp. N174
1.6	-	mutant D40G	Streptomyces sp. N174
21.9	-	mutant E36N	Streptomyces sp. N174
29.1	-	mutant E36D	Streptomyces sp. N174
29.2	-	mutant E36A	Streptomyces sp. N174
37.6	-	mutant T45S	Streptomyces sp. N174
44.7	-	mutant E36Q	Streptomyces sp. N174
52.9	-	wild-type	Streptomyces sp. N174

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
chitosan + H2O	catalyzes the hydrolysis of beta-1,4-glycosidic links	Streptomyces sp. N174	chitosan-oligosaccharides	-	?

Synonyms

Synonyms	Comment	Organism
CsnN174	-	Streptomyces sp. N174

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.0167	-	chitosan	mutant D40G/T45D, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
0.024	-	chitosan	mutant E36A/D40G, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
0.0368	-	chitosan	mutant T45E, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
0.0683	-	chitosan	mutant E36Q/D40G, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
0.44	-	chitosan	mutant D40G, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
6.827	-	chitosan	mutant E36N, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
8.34	-	chitosan	mutant E36D, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
10.48	-	chitosan	mutant T45S, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
11.47	-	chitosan	mutant E36A, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
11.66	-	chitosan	mutant E36Q, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174
12.4	-	chitosan	wild-type, assayed as the amount of D-glucosamine released from chitosan	Streptomyces sp. N174

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Release 2023.1 (January 2023)